Adoptively Transferring Of CD8α~- Dendritic Cells From Schistosoma Japonicum Infected Mice Inhibites OVA-induced Asthma By IL-10

ObjectiveTo explore which subset of dendritic cells(DC) from Schistosoma japonicum(SJ) infected mice played a critical role in inhibition of OVA-induced allergic asthma and its mechanism.Method 10 normal mice and 10 SJ infected mice were prepared for the extraction of CD8α+DC and CD8α-DC by MACS. We expressed the four DC subsets as NCD8α+DC,NCD8α-DC,SJCD8α+DC and SJCD8α-DC.Another 24 BALB/c mice were randomly divided into 4 groups,that is,SJCD8α+ group,SJCD8α- group,Asthma group,and Normal group. SJCD8α+DC and SJCD8α-DC were respectively transferred to SJCD8α+ group and SJCD8α- group. Then,allergic asthma was induced by OVA in SJCD8α+ group,SJCD8α- group and Asthma group simultaneously. Normal group was used as control.1. Histological analysis of the 4 groups were done by H&E staining.2. Total RNA of 4 DC subsets were extracted and were reversely transcribed to c DNA. Then IL-12 and IL-10 m RNA production were detected by q-PCR.3. Bronchial alveolar lavage fluid(BALF)from different groups were collected to determine IL-10 protein levels of them by ELISA.4. Spleen single-cell suspension from the 4 groups were prepared and spleen cells were cultured with OVA stimulation. 72 h later,spleen cell supernatant(SCS)were collected to determine IL-10 protein levels of them by ELISA.Result 1. Compared to that of Asthma group,the lung inflammatory responses in SJCD8α+ group were more or less alleviated--Numbers of inflammatory cells infiltrated in mesenchyme,mucous filled bronchial lumens,but bronchial walls were intact and not thickened so much. However,pathological changes of lungs from SJCD8α- group were slighter than those of Asthma group and SJCD8α+ group--Inflammatory infiltration was significantly reduced,bronchial walls were intact,not thickened and there was no mucous retention.2. IL-12 m RNA levels of NCD8α+DC、NCD8α-DC、SJCD8α+DC and SJCD8α-DC were 23.14±0.40, 18.74±2.92, 37.96±1.36 and 25.34±4.31(Expression/GAPDH %). IL-12 m RNA level of SJCD8α-DC was similar to that of NCD8α+DC and NCD8α-DC,and was lower than SJCD8α+DC(P<0.01). However,IL-10 m RNA level of NCD8α+DC、NCD8α-DC、SJCD8α+DC and SJCD8α-DC were 79.74±1.39、89.50±13.95、103.17±3.69 和 172.68±29.39. SJCD8α-DC showed higher IL-10 m RNA level than NCD8α-DC and SJCD8α+DC(P<0.05,P<0.05).3. IL-10 levels of SCS from Normal group,Asthma group,SJCD8α+ group and SJCD8α- group were( 142.23±11.96) pg/ml,( 164.42±26.47) pg/ml,(254.16±26.28)pg/ml 和(841.53±9.49)pg/ml. IL-10 production of SCS from SJCD8α- group showed higher IL-10 level than Normal group,Asthma group and SJCD8α+ group(P<0.001,P<0.001,P<0.001).4. IL-10 levels of BALF from Normal group,Asthma group,SJCD8α+ group and SJCD8α- group were( 151.63±10.40) pg/ml,( 144.54±10.18) pg/ml,(202.88±25.30) pg/ml and(411.21±6.38) pg/ml. IL-10 production of BALF from SJCD8α- group was higher than that of Normal group,Asthma group and SJCD8α+ group(P<0.001,P<0.001,,P<0.001).Conclusion 1. SJCD8α-DC played a critical role in inhibition of OVA-induced allergic asthma.-2. SJCD8αDC is tolerogenic DC.3. Adoptively transferring of SJCD8α-DC inhibits allergic asthma by increasing IL-10 production.